Matrix production in chondrocytes transfected with sex determining region Y-Box 9 and telomerase reverse transcriptase genes: an in vitro evaluation from monolayer culture to three-dimensional culture

Background:: This study aimed to observe the cartilaginous matrix production in SRY (sex determining region Y)-box 9 (SOX9)- and/or telomerase reverse transcriptase (TERT)-transfected chondrocytes from monolayer to three-dimensional (3D) culture. Methods:: The genes were transferred into chondrocyte...

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Main Authors:	Md Nazir, Noorhidayah, Zulkifly, Ahmad Hafiz, Khalid, Kamarul Ariffin, Zainol, Ismail, Zamli, Zaitunnatakhin, Sha'ban, Munirah
Format:	Article
Language:	English English English
Published:	Korean Tissue Engineering and Regenerative Medicine Society 2019
Subjects:	RA Public aspects of medicine
Online Access:	http://irep.iium.edu.my/79713/ http://irep.iium.edu.my/79713/ http://irep.iium.edu.my/79713/ http://irep.iium.edu.my/79713/1/79713_Matrix%20Production%20in%20Chondrocytes_MYRA.pdf http://irep.iium.edu.my/79713/2/79713_Matrix%20Production%20in%20Chondrocytes_SCOPUS.pdf http://irep.iium.edu.my/79713/3/79713_Matrix%20Production%20in%20Chondrocytes_WOS.pdf

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spelling	iium-797132020-03-20T14:37:52Z http://irep.iium.edu.my/79713/ Matrix production in chondrocytes transfected with sex determining region Y-Box 9 and telomerase reverse transcriptase genes: an in vitro evaluation from monolayer culture to three-dimensional culture Md Nazir, Noorhidayah Zulkifly, Ahmad Hafiz Khalid, Kamarul Ariffin Zainol, Ismail Zamli, Zaitunnatakhin Sha'ban, Munirah RA Public aspects of medicine Background:: This study aimed to observe the cartilaginous matrix production in SRY (sex determining region Y)-box 9 (SOX9)- and/or telomerase reverse transcriptase (TERT)-transfected chondrocytes from monolayer to three-dimensional (3D) culture. Methods:: The genes were transferred into chondrocytes at passage-1 (P1) via lipofection. The post-transfected chondrocytes (SOX9-, TERT- and SOX9/TERT) were analysed at P1, P2 and P3. The non-transfected group was used as control. The 3D culture was established using the chondrocytes seeded in a disc-shaped PLGA/fibrin and PLGA scaffolds. The resulting 3D “cells-scaffolds” constructs were analysed at week-1, -2 and -3. The histoarchitecture was evaluated using haematoxylin and eosin, alcian blue and safranin o stains. The quantitative sulphated glycosaminoglycan (sGAG) content was measured using biochemical assay. The cartilage-specific markers expression were analysed via real-time polymerase chain reaction. Results:: All monolayer cultured chondrocytes showed flattened, fibroblast-like appearance throughout passages. Proteoglycan and sGAG were not detected at the pericellular matrix region of the chondrocytes. The sGAG content assay indicated the matrix production depletion in the culture. The cartilage-specific markers, COL2A1 and ACAN, were downregulated. However, the dedifferentiation marker, COL1A1 was upregulated. In 3D “cells-scaffolds” constructs, regardless of transfection groups, chondrocytes seeded in PLGA/fibrin showed a more uniform distribution and produced denser matrix than the PLGA group especially at week-3. Both sGAG and proteoglycan were clearly visualised in the constructs, supported by the increment of sGAG content, quantitatively. Both COL2A1 and ACAN were upregulated in SOX9/TERT-PLGA and SOX9/TERT-PLGA/fibrin respectively. While, COL1A1 was downregulated in SOX9/TERT-PLGA. Conclusion:: These findings indicated that the SOX9/TERT-transfected chondrocytes incorporation into 3D scaffolds facilitates the cartilage regeneration which is viable structurally and functionally. © 2019, The Korean Tissue Engineering and Regenerative Medicine Society. Korean Tissue Engineering and Regenerative Medicine Society 2019-06-01 Article PeerReviewed application/pdf en http://irep.iium.edu.my/79713/1/79713_Matrix%20Production%20in%20Chondrocytes_MYRA.pdf application/pdf en http://irep.iium.edu.my/79713/2/79713_Matrix%20Production%20in%20Chondrocytes_SCOPUS.pdf application/pdf en http://irep.iium.edu.my/79713/3/79713_Matrix%20Production%20in%20Chondrocytes_WOS.pdf Md Nazir, Noorhidayah and Zulkifly, Ahmad Hafiz and Khalid, Kamarul Ariffin and Zainol, Ismail and Zamli, Zaitunnatakhin and Sha'ban, Munirah (2019) Matrix production in chondrocytes transfected with sex determining region Y-Box 9 and telomerase reverse transcriptase genes: an in vitro evaluation from monolayer culture to three-dimensional culture. Tissue Engineering and Regenerative Medicine, 16 (3). pp. 285-299. ISSN 1738-2696 E-ISSN 2212-5469 https://link.springer.com/article/10.1007/s13770-019-00191-1 10.1007/s13770-019-00191-1
repository_type	Digital Repository
institution_category	Local University
institution	International Islamic University Malaysia
building	IIUM Repository
collection	Online Access
language	English English English
topic	RA Public aspects of medicine
spellingShingle	RA Public aspects of medicine Md Nazir, Noorhidayah Zulkifly, Ahmad Hafiz Khalid, Kamarul Ariffin Zainol, Ismail Zamli, Zaitunnatakhin Sha'ban, Munirah Matrix production in chondrocytes transfected with sex determining region Y-Box 9 and telomerase reverse transcriptase genes: an in vitro evaluation from monolayer culture to three-dimensional culture
description	Background:: This study aimed to observe the cartilaginous matrix production in SRY (sex determining region Y)-box 9 (SOX9)- and/or telomerase reverse transcriptase (TERT)-transfected chondrocytes from monolayer to three-dimensional (3D) culture. Methods:: The genes were transferred into chondrocytes at passage-1 (P1) via lipofection. The post-transfected chondrocytes (SOX9-, TERT- and SOX9/TERT) were analysed at P1, P2 and P3. The non-transfected group was used as control. The 3D culture was established using the chondrocytes seeded in a disc-shaped PLGA/fibrin and PLGA scaffolds. The resulting 3D “cells-scaffolds” constructs were analysed at week-1, -2 and -3. The histoarchitecture was evaluated using haematoxylin and eosin, alcian blue and safranin o stains. The quantitative sulphated glycosaminoglycan (sGAG) content was measured using biochemical assay. The cartilage-specific markers expression were analysed via real-time polymerase chain reaction. Results:: All monolayer cultured chondrocytes showed flattened, fibroblast-like appearance throughout passages. Proteoglycan and sGAG were not detected at the pericellular matrix region of the chondrocytes. The sGAG content assay indicated the matrix production depletion in the culture. The cartilage-specific markers, COL2A1 and ACAN, were downregulated. However, the dedifferentiation marker, COL1A1 was upregulated. In 3D “cells-scaffolds” constructs, regardless of transfection groups, chondrocytes seeded in PLGA/fibrin showed a more uniform distribution and produced denser matrix than the PLGA group especially at week-3. Both sGAG and proteoglycan were clearly visualised in the constructs, supported by the increment of sGAG content, quantitatively. Both COL2A1 and ACAN were upregulated in SOX9/TERT-PLGA and SOX9/TERT-PLGA/fibrin respectively. While, COL1A1 was downregulated in SOX9/TERT-PLGA. Conclusion:: These findings indicated that the SOX9/TERT-transfected chondrocytes incorporation into 3D scaffolds facilitates the cartilage regeneration which is viable structurally and functionally. © 2019, The Korean Tissue Engineering and Regenerative Medicine Society.
format	Article
author	Md Nazir, Noorhidayah Zulkifly, Ahmad Hafiz Khalid, Kamarul Ariffin Zainol, Ismail Zamli, Zaitunnatakhin Sha'ban, Munirah
author_facet	Md Nazir, Noorhidayah Zulkifly, Ahmad Hafiz Khalid, Kamarul Ariffin Zainol, Ismail Zamli, Zaitunnatakhin Sha'ban, Munirah
author_sort	Md Nazir, Noorhidayah
title	Matrix production in chondrocytes transfected with sex determining region Y-Box 9 and telomerase reverse transcriptase genes: an in vitro evaluation from monolayer culture to three-dimensional culture
title_short	Matrix production in chondrocytes transfected with sex determining region Y-Box 9 and telomerase reverse transcriptase genes: an in vitro evaluation from monolayer culture to three-dimensional culture
title_full	Matrix production in chondrocytes transfected with sex determining region Y-Box 9 and telomerase reverse transcriptase genes: an in vitro evaluation from monolayer culture to three-dimensional culture
title_fullStr	Matrix production in chondrocytes transfected with sex determining region Y-Box 9 and telomerase reverse transcriptase genes: an in vitro evaluation from monolayer culture to three-dimensional culture
title_full_unstemmed	Matrix production in chondrocytes transfected with sex determining region Y-Box 9 and telomerase reverse transcriptase genes: an in vitro evaluation from monolayer culture to three-dimensional culture
title_sort	matrix production in chondrocytes transfected with sex determining region y-box 9 and telomerase reverse transcriptase genes: an in vitro evaluation from monolayer culture to three-dimensional culture
publisher	Korean Tissue Engineering and Regenerative Medicine Society
publishDate	2019
url	http://irep.iium.edu.my/79713/ http://irep.iium.edu.my/79713/ http://irep.iium.edu.my/79713/ http://irep.iium.edu.my/79713/1/79713_Matrix%20Production%20in%20Chondrocytes_MYRA.pdf http://irep.iium.edu.my/79713/2/79713_Matrix%20Production%20in%20Chondrocytes_SCOPUS.pdf http://irep.iium.edu.my/79713/3/79713_Matrix%20Production%20in%20Chondrocytes_WOS.pdf
first_indexed	2023-09-18T21:51:43Z
last_indexed	2023-09-18T21:51:43Z
_version_	1777413812954071040

Matrix production in chondrocytes transfected with sex determining region Y-Box 9 and telomerase reverse transcriptase genes: an in vitro evaluation from monolayer culture to three-dimensional culture

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